Differentiation of the plasma membrane of hepatic cells in monolayer cultures

Hepatocytes from rats were isolated by treatment with trypsin and cultured. Plasma membranes at different culture stages were observed by electron microscopy. The activities of 5' nucleotidase and adenosinetriphosphatase on the plasma membranes were examined. The cell coat was also studied by use of the concanavalin A-peroxidase technique. The surfaces of single cells, covered with microvilli, are the site of adenosinetriphosphatase activity only and are devoid of 5'-nucleotidase activity. After a few h of culture, the cells are grouped together in tight clusters or long trails and are separated by an intercellular space of 250 A, partially permeable to lanthanum nitrate. The juxtaposed plasma membranes on which 5'-nucleotidase and adenosinetriphosphatase activities occur also delimit spaces similar to bile canaliculi. The formation of junction complexes and their permeability to lanthanum nitrate was also studied. No enzymatic activity is observed at the junctions. The numerous tight junctions, impervious to the tracer, are always accompanied by a profusion of microfilaments. Mature desmosomes are rare, and are present only in the form of "maculae adhaerentes diminutae." The gap junctions, nearly always permeable to the tracer, form rapidly and assume a variety of shapes (trail, bulge and ring-like), the significance of which is open to discussion. The use of concanavalin A permits localization of the free sugar sites on the surface of the cells, in the pinocytotic vesicles and in the internal space of the gap junctions.     

LRG‐1 promotes fat graft survival through the RAB31‐mediated inhibition of hypoxia‐induced apoptosis

Autologous adipose tissue is an ideal soft tissue filling material, and its biocompatibility is better than that of artificial tissue substitutes, foreign bodies and heterogeneous materials. Although autologous fat transplantation has many advantages, the low retention rate of adipose tissue limits its clinical application. Here, we identified a secretory glycoprotein, leucine‐rich‐alpha‐2‐glycoprotein 1 (LRG‐1), that could promote fat graft survival through RAB31‐mediated inhibition of hypoxia‐induced apoptosis. We showed that LRG‐1 injection significantly increased the maintenance of fat volume and weight compared with the control. In addition, higher fat integrity, more viable adipocytes and fewer apoptotic cells were observed in the LRG‐1‐treated groups. Furthermore, we discovered that LRG‐1 could reduce the ADSC apoptosis induced by hypoxic conditions. The mechanism underlying the LRG‐1‐mediated suppression of the ADSC apoptosis induced by hypoxia was mediated by the upregulation of RAB31 expression. Using LRG‐1 for fat grafts may prove to be clinically successful for increasing the retention rate of transplanted fat.     

An Imported Case of Cystic Echinococcosis in the Liver

A 25-year-old Uzbek male presented with right upper abdominal pain for 20 days. On radiologic studies, a huge cystic mass was noticed in the right liver which was suspected as parasitic. The patient received right hepatic segmentectomy (segment 7), and the surgically resected mass was confirmed as cystic echinococcosis (CE), measuring 10.5 cm in its diameter. The inner surface of the cyst was bile-stained. The patient was discharged on the 8th hospital day, and was rechecked 6 months after the surgical intervention without any evidence of recurrence. The present report describes findings of an imported case of CE which represented ultrasound images of the ''ball of wool''.     

Gene expression profile in immunologically injured liver cell of mice

To study the gene expression profiles between immunologically injured liver cell and normal liver cell of mice and to screen on a large scale the differentially expressed genes associated with the formation of liver injury, the experimental mice were randomly divided into the normal group for controlling and the immunologically liver-injured group induced by BCG and LPS. The liver mRNA of the two groups were extracted respectively and reversely-transcribed to cDNA with the incorporation of different fluorescence (Cy3, Cy5) labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray chips. The fluorescent signal results were acquired by scanner ScanArray 4000 and analyzed with software GenePix Pro 3.0. Among the 14112 target genes, 293 genes were found to be significantly differentially expressed, in which 188 genes were up-regulated and 105 genes were down-regulated. Based on the analysis of biological functions of those differentially expressed genes, it was indicated that the occurrence and development of mouse liver damage induced by BCG and LPS were highly correlated with the processes of immune reactions, cell synthesis, metabolism, apoptosis and transportation in liver cell, which might be quite important for elucidating the regulatory network of gene expression associated with the liver damage, also important for finally discovering the pathogenic mechanisms of immunological liver damage.     

小鼠胎肝细胞透明质酸3D培养体系的建立

目的 利用透明质酸建立小鼠胎肝细胞3D培养体系。 方法 分离获得胚胎12-14天胎肝细胞,利用KM培养基进行初步2D肝干/祖细胞的筛选培养,并利用透明质酸及KM培养基配制水凝胶建立3D细胞培养体系。 结果 胎肝细胞在2D体系中呈现克隆状生长。分离培养获得的肝干/祖细胞克隆在透明质酸建立的3D培养体系保持增殖活性,并进一步获得肝细胞功能特性,表现为3D培养上清中白蛋白合成和尿素水平显著增加。Q-PCR结果显示随着3D培养时间的延长,其肝细胞干性标志如AFP、CK19、EpCAM、Prox1等表达水平都大幅度降低且接近成年小鼠肝脏表达水平。 结论 本研究成功建立基于透明质酸的小鼠胎肝细胞的3D无血清培养体系,并可促进小鼠胎肝细胞肝细胞功能进一步成熟。     

原发性肝癌患者术后癌因性疲乏的影响因素及其预测模型构建

摘要 目的：分析原发性肝癌患者术后癌因性疲乏（CRF）的影响因素并构建预测模型。方法：选取2020年1月～2023年1月湖南师范大学附属第一医院收治接受手术治疗的200例原发性肝癌患者,根据术后3个月是否存在CRF将患者分为CRF组（124例）和非CRF组（76例）。单因素和多因素Logistic回归分析影响原发性肝癌患者术后CRF的因素并构建其预测模型。通过受试者工作特征（ROC）曲线分析预测模型对原发性肝癌患者术后CRF的预测价值。结果：单因素分析显示,CRF组病程长于非CRF组,Child-Pugh分级B级、美国东部肿瘤协作组功能状态（ECOG）评分1～2分、辅助化疗、医疗付费方式自费、抑郁/焦虑比例高于非CRF组,文化程度高中及以上、家庭月收入＞3000元、高度社会支持度比例低于非CRF组（P＜0.05）。多因素Logistic回归分析显示,病程延长、Child-Pugh分级B级、ECOG评分1～2分、辅助化疗、医疗付费方式自费、抑郁/焦虑为影响原发性肝癌患者术后CRF的独立危险因素,家庭月收入＞3000元、高度社会支持为独立保护因素（P＜0.05）。原发性肝癌患者术后CRF的预测模型方程：Logit（P）=P/1-P=-1.252+0.409×病程+0.839×Child-Pugh分级+1.378×ECOG评分+1.055×辅助化疗+1.476×医疗付费方式-0.793×家庭月收入+0.883×抑郁/焦虑-1.260×社会支持度。霍斯默-莱梅肖检验P＞0.05。ROC曲线分析显示,模型预测原发性肝癌患者术后CRF的曲线下面积为0.910,敏感度为87.10%,特异度为85.53%。结论：病程、Child-Pugh分级、ECOG评分、辅助化疗、医疗付费方式、抑郁/焦虑、家庭月收入、社会支持度为影响原发性肝癌患者术后CRF的因素,基于此构建的预测模型对原发性肝癌患者术后CRF的预测价值较高,可能有助于临床早期发现和干预原发性肝癌患者术后CRF,以改善患者预后。     

Adaptive changes in lipid composition of rat liver plasma membrane during postnatal development following maternal ethanol ingestion

The fatty acid composition of constituent phospholipids and the cholesterol content of rat liver plasma membranes were determined subsequent to maternal alcohol ingestion during pregnancy and lactation. The alcoholic group was given a liquid Metrecal diet containing 37% ethanol-derived calories. The control group was pair-fed an isocaloric sucrose/Metrecal diet. Litters were killed for lipid analyses at days 5, 15 and 25 after birth. These studies revealed that the total phospholipid phosphorus was similar and increased significantly with age in both groups. Cholesterol also increased significantly with age in both groups but was greater in the alcoholic pups, resulting in a higher cholesterol/phospholipid molar ratio. While the phosphatidylethanolamine (PE) content increased with age in both groups, that of sphingomyelin decreased. Phosphatidylserine + phosphatidylinositol (PS + PI) was significantly higher in the control group at all ages studied. A consistent increase of C22:6 in phosphatidylcholine (PC), sphingomyelin, PS + PI and in the total phospholipid fraction from alcoholic pups was observed. Although other fatty acid changes were found in PC, PS + PI and sphingomyelin, PE was not affected. These results suggest that specific adaptive changes were induced in the liver plasma membrane lipids of the progeny from alcoholic rats.